Isolation of Cryptosporidium Oocysts and Sporozoites Using Discontinuous Sucrose and Isopycnic Percoll Gradients

Arrowood, Michael J.; Sterling, Charles R.

doi:10.2307/3282084

Cited by 385 publications

(200 citation statements)

References 13 publications

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“…5% potassium dichromate solution until they were used. The oocysts were purified by a sucrose and Percoll gradient method [22]. DNA was isolated from the purified oocysts as previously described [23].…”

Section: Genotypingmentioning

confidence: 99%

Genotyping of Cryptosporidium spp. isolated from human stool samples in Switzerland

et al. 2003

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SUMMARYIn a study to estimate the frequency of Cryptosporidium infections in Switzerland, stool samples from patients found to be positive for Cryptosporidium spp. by modified Ziehl-Neelson staining and fluorescence microscopy were used for genotyping experiments. With 9 of 12 samples, DNA extraction and subsequent genotyping was successful. All Cryptosporidium-isolates belonged to the bovine genotype. In one stool sample, two strains of Cryptosporidium were demonstrated, suggesting a mixed infection. In comparison with reference strains from calves, one of the isolates showed a full sequence identity and the other a similarity of 97 . 5%. The fact that only bovine genotypes were detected suggests, that cryptosporidiosis must primarily be considered as a zoonotic disease in Switzerland. This is in contrast to other countries, where the human genotype of C. parvum was shown to dominate the epidemiological situation. The results of our study are supported by the previous finding, that two of the analysed strains originated from patients who used to consume raw milk or raw cream, a known risk factor for cryptosporidiosis.

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Section: Genotypingmentioning

confidence: 99%

Genotyping of Cryptosporidium spp. isolated from human stool samples in Switzerland

et al. 2003

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“…The feces diluted about 1:5 in sucrose solution (SG: 1.27) and centrifuged at 2,0009g for 15 min. After centrifugation, the supernatant containing oocyst discarded and collected (Arrowood and Sterling 1987).…”

Section: Oocyst Preparation and Purificationmentioning

confidence: 99%

Immune response of newborn BALB/c mice to Cryptosporidium infection

et al. 2014

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Cryptosporidium parvum is a protozoan parasite which causes diarrheal in human and animals worldwide. Infection transmission has reported through oralfecal by infectious objects through foods and drinks. In this study we explored the immune response pathway in animal model for C. parvum to develop the new treatment way. Oocysts collected from fecal positive for C. parvum and diluted about 1:5 in sucrose solution. New born BALB/c mice (3 days) divided to 2 different groups. Control group hadn't received any oocyst, the test groups received 5 9 10 5 oocysts. 5 mice selected for each control group and 11 mice chosen for each test group. Blood collected from heart bleeds in days of 6, 9, 12 and 16. Protein concentrations determined by bio-photometer. Dot blotting used to find out total antibody concentrations oocyst antigen. Among the test and the control groups, blots appeared in test group which means antibody production, but not any blot observed in the control groups. The non-characteristic proteins in serum were measured by the biophotometer. In this study, we investigated antibody serum production against C. parvum oocysts in new born BALB/c mice. The detected antibody through dot blot technique was our aims which had conjugated to our characteristic antiserum. The recorded numbers for the controls by biophotometer related to the non characteristic proteins in serum. The results of this study can used to produce polyclonal or monoclonal antibodies against cryptosporidiosis.

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“…The feces were collected daily, mixed with equal volumes of 2.5% potassium dichromate (K2Cr2O7), and stored at 4℃. Oocysts were isolated by means of discontinuous sucrose gradients [14]. Briefly, Sheather's solution containing sucrose was diluted with 0.025 M phosphate-buffered saline (PBS) supplemented with 1% Tween 80 to make 1 : 2 and 1 : 4 gradient solutions.…”

Section: Antigen Preparationmentioning

confidence: 99%

Antibody Responses to Cryptosporidium Antigen in HIV-positive Patients in the Republic of Korea

et al. 2008

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Abstract:The diagnosis of cryptosporidiosis has been carried out using coprologic techniques in the Republic of Korea. However, antibody responses to Cryptosporidium have rarely been studied. Serum antibodies from HIV-positive/oocyst-positive Korean patients recognized significantly 31 and 27 kDa antigens, and HIV-negative/oocyst-positive individuals clearly reacted to 15/17 kDa antigens. Compared with oocyst-positive cases, 18.7% and 75.8% of sera from HIV-positive patients reacted to 31 and 27 kDa antigens. Only 11.1% of HIV-negative individuals reacted to 15/17 kDa. Based on these findings, serum antibody responses were different between HIV-positive and HIV-negative individuals infected with Cryptosporidium, and it is suggested that HIV-positive patients are more frequently exposed to C. parvum compared to HIV-negative individuals.

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Isolation of Cryptosporidium Oocysts and Sporozoites Using Discontinuous Sucrose and Isopycnic Percoll Gradients

Cited by 385 publications

References 13 publications

Genotyping of Cryptosporidium spp. isolated from human stool samples in Switzerland

Genotyping of Cryptosporidium spp. isolated from human stool samples in Switzerland

Immune response of newborn BALB/c mice to Cryptosporidium infection

Antibody Responses to Cryptosporidium Antigen in HIV-positive Patients in the Republic of Korea

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